Small Interfering RNA Targeting Connective Tissue Growth Factor Interference In The Reverse Impact Of Rats Liver Fibrosis

Objectives: This research by design and the synthesis of the liver fibrosis key gene layer of small interfering RNA targeting connective tissue growth factor,and through cell transfection bloting high-performance CTGF siRNA of liver fibrosis sequence. To explore effect of small interfering RNA targeting connective tissue growth factor on liver function and fibre index of rats with liver fibrosis and whether inhibit CTGF expression in rats liver in vivo and prevent rats hepatic fibrosis.Methods: 1.Reference siRNA design principles, the application of RNA design software online, the design of RNA interference three candidates, and to transfection their normal cell(L-02),transfection non-specificity siRNA(non-homology of CTGF mRNA) in comparison,with western blot detection L-02 cell CTGF protein, expressed by the albumen bloting the interference sequence of CTGF.2. Thirty male rats were randomly divided into five groups,Rats received intraperitoneally injection of 40% CCl4(3 ml/kg) together with tail vein injection of saline every three days for 8 consecutive weeks were served as model group;CCl4 together with tail vein delivery of siRNA (0.1 mg/kg) as preventive group; CCl4 for 2 weeks followed by CCl4 and CTGF siRNA for more than 6 weeks as curative group; CCl4 for 4 weeks followed by CCl4 and CTGF siRNA for more than 4 weeks as advanced curative group and only tail vein injection of saline as control group. Tail vein pressure in all rats at 3 days af ter the last CCl4 injection were blood and hepatic tissue from rats were harvested. Serum transaminase(ALT,AST), albumin(ALB), total bilirubin(TBIL) and hepatic fibrosis indices were measured. Expression of CTGF mRNA and protein in rats liver was evaluated by RT-PCR and Western blot, respectively. Inflammation and fibrosis in rats liver was analyzed by H-E.Results:1. Compared with the contrast, the transfection siRNA the albumen L-02 CTGFexpress down regulation,and siCTGF-1,si-CTGF-2,siCTG-3 ratio of interference for 44.91%,93.99%,81.34%,with effect from the highest rate of the most obvious. To transfection nonspecific siRNA L-02 of the albumen cell CTGF expressed no marked change(p<0.05).2. Compared with model group, the expression of CTGF mRNA and protein in liver in both preventive and curative groups were markedly down-regulated (p<0.05). Inflammation, necrosis and fibrosis in hepatic tissue was significantly attenuated. In addition, the serum concent ration of transaminases liver fibrosis indices were greatly reduced.Compared with model group, the expression of CTGF mRNA and protein in liver in advanced curative group were down-regulated(p<0.05). Inflammation, necrosis and fibrosis in hepatic tissue was reduced. The serum concent ration of transaminases liver fibrosis indices were reduced. Compared with preventive and curative groups, the expression of CTGF mRNA and protein in liver in advanced curative group were up-regulated (p<0.05). Inflammation, necrosis and fibrosis in hepatic tissue was relative increased. The serum concent ration of transaminases liver fibrosis indices were relatively higher.Conclusion:Different CTGF siRNA L-02 CTGF expression to the cell level of efficiency of different and synthesis could be efficient to dampen these CTGF express CTGF siRNA. Tail vein delivery of CTGF siRNA significantly inhibit CTGF expression in rat s liver in vivo, and effectively prevent rat s hepatic fibrosis, Injection earlier, the more effectively prevent the progress of liver fibrosis, and liver fibrosis in advanced for a therapeutic effect, also suggesting siRNA targeting CTGF treatment of liver fibrosis will become a new taruet.